Development of an osteoblast-based 3D continuous-perfusion microfluidic system for drug screening |
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Authors: | Kihoon Jang Kae Sato Kazuyo Igawa Ung-il Chung Takehiko Kitamori |
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Institution: | (1) Department of Applied Chemistry, Graduate School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo, Tokyo 113-8656, Japan;(2) Core Research for Evolutional Science and Technology, Japan Science and Technology Agency, 4-1-8 Honcho, Kawaguchi, Saitama 332-0012, Japan;(3) Center for NanoBio Integration, The University of Tokyo, 7-3-1 Hongo, Bunkyo, Tokyo 113-8656, Japan;(4) Division of Tissue Engineering, The University of Tokyo Hospital, 7-3-1 Hongo Bunkyo-ku, Tokyo 113-8655, Japan;(5) Department of Bioengineering, School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo, Tokyo 113-8656, Japan |
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Abstract: | In this work, we demonstrated that biological cells could be cultured in a continuous-perfusion glass microchip system for
drug screening. We used mouse Col1a1GFP MC-3T3 E1 osteoblastic cells, which have a marker gene system expressing green fluorescent
protein (GFP) under the control of osteoblast-specific promoters. With our microchip-based cell culture system, we realized
automated long-term monitoring of cells and sampling of the culture supernatant system for osteoblast differentiation assay
using a small number of cells. The system successfully monitored cells for 10 days. Under the 3D microchannel condition, shear
stress (0.07 dyne/cm2 at a flow rate of 0.2 μL/min) was applied to the cells and it enhanced the GFP expression and differentiation of the osteoblasts.
Analysis of alkaline phosphatase (ALP), which is an enzyme marker of osteoblasts, supported the results of GFP expression.
In the case of differentiation medium containing bone morphogenetic protein 2, we found that ALP activity in the culture supernatant
was enhanced 10 times in the microchannel compared with the static condition in 48-well dishes. A combined system of a microchip
and a cell-based sensor might allow us to monitor osteogenic differentiation easily, precisely, and noninvasively. Our system
can be applied in high-throughput drug screening assay for discovering osteogenic compounds. |
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Keywords: | Bioassays Cell systems Microfluidics Osteoblast Shear stress Differentiation |
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